During the previous funding cycle, our program project entitled ?Staphylococcal biofilm and disease? has employed in-depth mechanistic approaches to define the developmental and metabolic processes important in Staphylococcus aureus biofilm development. A key aspect of the synergy of this project is that the knowledge gained was used as context to provide a more detailed understanding of the acquisition of available nutrients within specific host niches, as well as the impact of biofilm growth on the host immune response. These studies have led to a greatly enhanced understanding of the way in which S. aureus adapts to a host environment, providing new fundamental insight into biofilm development and novel approaches to the clinical management of staphylococcal disease. The overall hypothesis driving the goals of the proposed program project, S. aureus biofilm development creates unique metabolic niches that promote an immune suppressive environment, is a natural outgrowth of the current funding cycle and is tested in four synergistic and complementary projects that encompass a broad spectrum of synergistic and highly collaborative activities ranging from the basic biology of biofilm development and matrix regulation, to the host-associated metabolic processes that influence the immune response. To support the efforts of these four projects, we propose a continuation of our Bioimaging Core that maintains a BioFlux microfluidics system for biofilm growth and analysis, confocal microscopy, and an In Vivo Imaging System (IVIS). In addition, we propose a new Metabolomics Core that will establish and maintain the protocols and modeling needed to support the four projects associated with this PPG. Importantly, our vision is that the work of this core will lead to the development of a web-based metabolomics tool (funded through a separate mechanism) that will serve not only as an education tool to enhance the overall understanding of the S. aureus metabolome, but also as a hypothesis generator in support of scientific inquiry. Once this tool is established and validated, we will then make it available to the entire staphylococcal research community as a web-based resource that is integrated with our existing Nebraska Transposon Mutant Library (NTML) website. Finally, we propose an Administrative Core that will provide the administrative support needed to maximize the interactions between project leaders and to ensure that their projects maintain optimal synergy.